Progressive degeneration of nigrostriatal (NS) dopamine (DA) neurons underlies the motor symptoms of Parkinson's disease (PD) and therapies that slow progression are lacking. Abnormal DA metabolism has been proposed as a mechanism for the selective degeneration of NSDA neurons. All DA neurons, however, are not affected to the same extent in PD. While there is severe loss of midbrain NSDA neurons, the hypothalamic tuberoinfundibular (TI) DA neurons remain intact. A similar pattern of susceptibility can be seen in these DA neuronal populations when they are exposed to mitochondrial complex I inhibitors in vitro and in vivo. Although NSDA and TIDA neurons have a similar initial response to complex I inhibition, only TIDA neurons are able to recover using a mechanism that is dependent on new protein synthesis. Our preliminary data also implicate that parkin is involved in this differential susceptibility since it is markedly upregulated in TIDA, but not NSDA neurons following an acute toxic injury. Parkin is a multifunctional protein that supports normal mitochondrial function, has an E3 ligase that tags misfolded proteins for proteosomal degradation and appears to protect DA neurons from a variety of toxic insults. Parkin also has important proteosome independent functions that could promote DA neuronal survival. The overall goal of this project is to elucidate the parkin-mediated molecular mechanisms that render NSDA neurons sensitive and TIDA neurons resistant to exogenous neurotoxin exposure. We hypothesize that parkin, acting primarily via proteosome-independent mechanisms, protects TIDA neurons and can rescue NSDA neurons from injury induced by a PD relevant pathological stress, mitochondrial complex I inhibition. To address the central hypothesis, rAAV-mediated, spatially restricted manipulation of parkin expression will be performed in wild-type mice and on mice with a parkin null background. The impact of exogenously modulating parkin expression on the TIDA and NSDA neuronal responses to both acute and chronic neurotoxic stress will be assessed to confirm the central role of parkin in allowing DA neurons to recover following an initial insult. Molecular and pharmacological methods will be used to elucidate the initial downstream mediators of the neuroprotective effects of parkin. Despite the observation that a significant portion of DA neurons have been lost at the onset of clinically observable symptoms of PD, it is plausible that there are neurons that are at risk and dysfunctional, but that could recover if provided the necessary machinery for recovery. Since TIDA neurons are unique amongst DA neurons in their ability to recover from an injury induced by a PD relevant pathological stressor, they provide a powerful platform to dissect the parkin-mediated mechanisms of DA neuronal recovery and could yield novel targets for neuroprotective therapy development. PUBLIC HEALTH RELEVANCE: Parkin protein appears to facilitate the recovery of neurons following exposure to pathological stressors seen in Parkinson's disease. The protective mechanisms of parkin will be determined as a strategy to develop new therapies for Parkinson's disease.